Complete Growth Inhibition Research Articles

Probiotic-derived ferrichrome induces DDIT3-mediated antitumor effects in esophageal cancer cells

Esophageal cancer, which is common among the elderly, has the poorest prognosis among gastrointestinal cancers. Previously, we demonstrated that ferrichrome, produced by the probiotic Lactobacillus casei, exhibited anti-tumor effects in various gastrointestinal cancers, including colorectal and gastric cancers, with minimal effects on non-cancerous intestinal cells. However, it remains unclear whether ferrichrome exerts anti-tumor effects in esophageal cancer. A sulforhodamine B assay revealed that ferrichrome suppressed esophageal adenocarcinoma (OE33, OE19) and squamous cell carcinoma (KYSE70) cells. Ki-67 staining indicated that ferrichrome inhibited the proliferation of esophageal cancer cells. Cell cycle analysis showed that ferrichrome inhibited the DNA synthesis. TUNEL staining revealed that ferrichrome-induced DNA fragmentation. We also confirmed the cleavage of caspase-9 and PARP in ferrichrome-treated cells. Reverse transcription polymerase chain reaction demonstrated an increase in the mRNA of DNA damage-inducible transcript 3 (DDIT-3), a key regulator of programmed cell death, in ferrichrome-treated OE33 cells. In an in vivo OE33 xenograft model, intraperitoneal administration of 5-mg/kg ferrichrome for 14 days resulted in an almost complete inhibition of tumor growth. However, 14 days of intraperitoneal administration of 20-mg/kg 5-fluorouracil (5-FU), but not 20-mg/kg ferrichrome, induced weight loss and myelosuppression in both young and aged mice. Our findings indicate that ferrichrome induces DNA damage-inducible transcript-3, thereby producing anti-tumor effects, including cell cycle arrest and apoptosis, with minimal adverse effects in esophageal cancer cells. This illustrates the high potential of ferrichrome as an anti-tumor drug against esophageal carcinoma.

In vitro effects of phytogenic feed additive on Piscirickettsia salmonis growth and biofilm formation.

Piscirickettsiosis is the main cause of mortality in salmonids of commercial importance in Chile, which is caused by Piscirickettsia salmonis, a Gram-negative, γ-proteobacteria that can produce biofilm as one of its virulence factors. The Chilean salmon industry uses large amounts of antibiotics to control piscirickettsiosis outbreaks, which has raised concern about its environmental impact and the potential to induce antibiotic resistance. Thus, the use of phytogenic feed additives (PFA) with antibacterial activity emerges as an interesting alternative to antimicrobials. Our study describes the antimicrobial action of an Andrographis paniculate-extracted PFA on P. salmonis planktonic growth and biofilm formation. We observed complete inhibition of planktonic and biofilm growth with 500 and 400 μg/mL of PFA for P. salmonis LF-89 and EM-90-like strains, respectively. Furthermore, 500 μg/mL of PFA was bactericidal for both evaluated bacterial strains. Sub-inhibitory doses of PFA increase the transcript levels of stress (groEL), biofilm (pslD), and efflux pump (acrB) genes for both P. salmonis strains in planktonic and sessile conditions. In conclusion, our results demonstrate the antibacterial effect of PFA against P. salmonis invitro, highlighting the potential of PFA as an alternative to control Piscirickettsiosis.

Management of Fusarium fruit rot of Luffa cylindrical caused by Fusarium incarnatum and Fusarium chlamydosporum

Fusarium rot of Luffa cylindrical was detected at Kafr-El-Dawar province, Beheira governorate, North Egypt during the growing season 2021. Surveyed disease incidence at Beheira, Kafr El-Shekh and Sharkia governorate showed fruit rot incidences of 18.33%, 16.12%, and 12.42%, respectively. Two fungal isolates of Fusarium spp. were tested for their pathogenic ability to induce Fusarium rot disease and typical disease symptoms. The online Basic Local Alignment Search Tool for Nucleotide Sequences (BLASTN) was used for identification and confirmation of identity performance of Internal Transcribed Spacer (ITS) and DNA sequencing, then compared with sequences available in GenBank. The two fungal isolates were identified as Fusarium incarnatum and Fusarium chlamydosporum and registered under accession number MN480497 and MN480498, respectively. Under laboratory conditions, organic acids, essential oils and organic salts significantly inhibited the growth of the two fungal isolates. Complete inhibition of fungal growth (100%) was recorded at 0.75 g/l of Topsin-M70 (Thiophanate-methyl) and 0.5% of salicylic acid. Meanwhile, a moderate growth reduction was observed using benzoic acid, coumarine, and cinnamon, thyme oil and garlic oil treatments. Under field conditions for two successive seasons, similar trends were observed. Foliar sprays with the fungicide Topsin-M70 showed the largest reduction in disease expression. The highest fruit rot disease reduction was recorded for salicylic acid and coumarin treatments followed by thyme oil, benzoic acid and potassium carbonate, garlic oil, boric acid and sodium bicarbonate. We concluded that the treatments are effective, safe and cost-effective methods for the control of this pre-harvest fruit disease under field conditions.

Versatile Antibacterial and Antioxidant Bacterial Cellulose@Nanoceria Biotextile: Application in Reusable Antimicrobial Face Masks.

Despite considerable interest in medical and pharmaceutical fields, there remains a notable absence of functional textiles that concurrently exhibit antibacterial and antioxidant properties. Herein, a new composite fabric constructed using nanostructured bacterial cellulose (BC) covalently-linked with cerium oxide nanoparticles (BC@CeO2NPs) is introduced. The synthesis of CeO2NPs on the BC is performed via a microwave-assisted, in situ chemical deposition technique, resulting in the formation of mixed valence Ce3+/Ce4+ CeO2NPs. This approach ensures the durability of the composite fabric subjected to multiple washing cycles. The Reactive oxygen species (ROS) scavenging activity of CeO2NPs and their rapid and efficient eradication of >99% model microbes, such as Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus remain unaltered in the composite. To demonstrate the feasibility of incorporating the fabric in marketable products, antimicrobial face masks are fabricated with filter layers made of BC@CeO2NPs cross-linked with propylene or cotton fibers. These masks exhibit complete inhibition of bacterial growth in the three bacterial strains, improved breathability compared to respirator masks and enhanced filtration efficiency compared to single-use surgical face masks. This study provides valuable insights into the development of functional BC@CeO2NPs biotextiles in which design can be extended to the fabrication of medical dressings and cosmetic products with combined antibiotic, antioxidant and anti-inflammatory activities.

Hypervirulent Capsular Serotypes K1 and K2 Klebsiella pneumoniae Strains Demonstrate Resistance to Serum Bactericidal Activity and Galleria mellonella Lethality.

Hypervirulent Klebsiella pneumoniae (hvKp) is a variant that has been increasingly linked to severe, life-threatening infections including pyogenic liver abscess and bloodstream infections. HvKps belonging to the capsular serotypes K1 and K2 have been reported worldwide, however, very scarce studies are available on their genomics and virulence. In the current study, we report four hypermucoviscous extended-spectrum β-lactamase-producing hvKp clinical strains of capsular serotype K1 and K2 isolated from pus and urine of critically ill patients in tertiary care hospitals in Oman. These strains belong to diverse sequence types (STs), namely ST-23(K1), ST-231(K2), ST-881(K2), and ST-14(K2). To study their virulence, a Galleria mellonella model and resistance to human serum killing were used. The G. mellonella model revealed that the K1/ST-23 isolate was the most virulent, as 50% of the larvae died in the first day, followed by isolate K2/ST-231 and K2/ST-14, for which 75% and 50% of the larvae died in the second day, respectively. Resistance to human serum killing showed there was complete inhibition of bacterial growth of all four isolates by the end of the first hour and up to the third hour. Whole genome sequencing (WGS) revealed that hvKp strains display a unique genetic arrangement of k-loci. Whole-genome single-nucleotide polymorphism-based phylogenetic analysis revealed that these hvKp isolates were phylogenetically distinct, belonging to diverse clades, and belonged to different STs in comparison to global isolates. For ST-23(K1), ST-231(K2), ST-881(K2), and ST-14(K2), there was a gradual decrease in the number of colonies up to the second to third hour, which indicates neutralization of bacterial cells by the serum components. However, this was followed by a sudden increase of bacterial growth, indicating possible resistance of bacteria against human serum bactericidal activity. This is the first report from Oman detailing the WGS of hvKp clinical isolates and assessing their resistance and virulence genomics, which reinforce our understanding of their epidemiology and dissemination in clinical settings.

Antifungal activity of essential oil against Botrytis cinerea in Vitis vinífera L

Grapes (Vitis vinífera L) is extensively cultivated and widely consumed fruit world wide But it is also very highly susceptible to fungal infections such as B cinérea which increases post-harvest losses, quality deterioration, and making it unsuitable for human consumption. This results in significant economic loss of grapes and impacts grape farmers’ profits. Therefore, there is an urgent need to identify potential methods and approaches to decrease post-harvest losses of grapes caused by B. cinérea. Currently, synthetic fungicides are used to control grape infections which has harmful impact on the environment and human health. Therefore, essential oil from plants sources could be a potential alternative to synthetic fungicides. In this study, antifungal activity of three essential oils was evaluated against B. cinérea using fumigation method. In an in vitro study, the essential oils extracted from Zingiber officinale and Saturaga montana, each at a concentration of 4mg/mL, exhibited complete inhibition (100%) of fungal growth. The physiological study of fruit in term of TSS, pH and weight loss showed that there is slow deterioration of fruit during storage. In-vivo results showed that essential oil from natural source can be potential alternative for synthetic fungicides which is ecofriendly and effective to control diseases and post-harvest losses in grapes.

Atividade antifúngica e mecanismo de ação de monoterpenos contra Botrytis cinerea

ABSTRACT Botrytis cinerea is considered one of the most important post-harvest pathogens being the causative agent of gray rot. To reduce the use of synthetic fungicides, it is important to explore alternative products with antifungal properties. Among these alternative products are essential oils, which present monoterpenes as major compounds. The objective of this study was to evaluate the effects of eight monoterpenes (1,8-cineole, carvacrol, citral, citronellal, citronellol, geraniol, linalool, and thymol) on the control of B. cinerea. The mycelial growth of B. cinerea was assessed after treating it with the monoterpenes at a concentration of 1,000 mg/L. Subsequently, the minimal inhibitory concentrations (IC90) of the monoterpenes that showed the greatest antifungal potential were determined. Carvacrol and thymol were tested on B. cinerea cell membrane integrity, intracellular accumulation of reactive oxygen species (ROS), and mitochondrial membrane potential of the conidia. Among the tested monoterpenes carvacrol, citral, citronellal, citronellol, geraniol, and thymol demonstrated complete inhibition of mycelial growth at a concentration of 1,000 mg/L. Carvacrol and thymol exhibited the lowest IC90 values against B. cinerea, with an IC90 of 125 mg/L. Furthermore, carvacrol and thymol induced conidial death in a dose-dependent manner, resulting in the disruption of cell membrane integrity, increased intracellular ROS levels, and decreased mitochondrial membrane potential. These findings highlight the potential of carvacrol and thymol as alternative means of controlling B. cinerea.

Silicon exhibits dose-dependent impact on barley growth

As population growth accelerates, agriculture’s significance in our lives remains paramount. However, this surge in population has led to land degradation and increased food scarcity. The impact of silicon (Si) on plants has garnered significant attention in agricultural research. This study was aimed at examining the impact of Si on barley (Hordeum vulgare) plants by administering varying concentrations (5µM, 10µM, 20µM, 50µM, 100µM, 200µM, 500µM, 1000µM) and assessing their effects on plant growth parameters, particularly root and shoot lengths, and root fresh weight. Our study employed a controlled experimental setup to observe how barley plants respond to varying concentrations of Si. The data indicates that Si concentrations up to 10µM offer advantageous effects on barley compared to the control group, suggesting its potential in bolstering agricultural productivity. Similarly, concentrations of 20µM, 50µM, and 100µM were identified as safe for plant growth, opening avenues for their agricultural utilization. However, concentrations of 500µM and 1000µM resulted in complete inhibition of plant growth, emphasizing the necessity for cautious Si application in agriculture to prevent adverse impacts on crop yields. These findings underscore the importance of fine-tuning Si concentrations in agricultural practices to optimize benefits while mitigating potential risks to plants.

Lactide oligomers modified with linear polyethyleneimine for antibacterial coatings

Antibacterial polylactide film composed of commercial PLA and block copolymer of lactide and linear polyethyleneimine (L-PEI) has been prepared. To observe complete inhibition of bacterial growth only 1.1 wt% of linear L-PEI is required.

Treatment with novel topoisomerase inhibitors in Ewing sarcoma models reveals heterogeneity of tumor response.

The topoisomerase 1 (TOP1) inhibitor irinotecan is a standard-of-care agent for relapsed Ewing sarcoma (EWS), but its efficacy is limited by chemical instability, rapid clearance and reversibility, and dose-limiting toxicities, such as diarrhea. Indenoisoquinolines (IIQs) represent a new class of clinical TOP1 inhibitors designed to address these limitations. In this study, we evaluated the preclinical efficacy of three IIQs (LMP400, LMP744, and LMP776) in relevant models of EWS. We characterized the pharmacokinetics of IIQs in orthotopic xenograft models of EWS, optimized the dosing regimen through tolerability studies, and tested the efficacy of IIQs in a panel of six molecularly heterogeneous EWS patient-derived xenograft (PDX) models. For each PDX, we conducted whole genome and RNA sequencing, and methylation analysis. We show that IIQs potently inhibit the proliferation of EWS cells in vitro, inducing complete cell growth inhibition at nanomolar concentrations via induction of DNA damage and apoptotic cell death. LMP400 treatment induced ≥30% tumor regression in two of six PDX models, with more durable regression compared to irinotecan treatment in one of these models. RNA sequencing of PDX models identified a candidate predictive biomarker gene signature for LMP400 response. These data, along with pharmacogenomic data on IIQs in sarcoma cell lines, are available at a new interactive public website: https://discover.nci.nih.gov/rsconnect/EwingSarcomaMinerCDB/. Our findings suggest that IIQs may be promising new agents for a subset of EWS patients.

Self-controllable proteinic antibacterial coating with bacteria-triggered antibiotic release for prevention of periprosthetic infection

Periprosthetic infection is a devastating postimplantation complication in which a biofilm layer harboring invasive microorganisms forms around orthopedic implants, leading to severe implant failure and patient morbidity. Despite the development of several infection-triggered antibiotic release approaches, most current antibacterial coatings are susceptible to undesired antibiotic leakage or mechanical disintegration during prosthesis installation. Herein, we propose a self-controllable proteinic antibacterial coating capable of both long-lasting adherence onto titanium implant substrates over the implant fixation period and instantaneous bacterial eradication. Importantly, the pH-dependent reversible metal coordination of mussel adhesive protein (MAP) enabled bacterial concentration-dependent antibiotic delivery in response to infection-induced acidification. In addition, the MAP coating exhibited superior self-healable adhesive properties and scratch resistance, which enabled to avert issues associated with mechanical damages, including peeling and cracking, often occurring in conventional implant coating systems. The gentamicin-loaded MAP coating exhibited complete inhibition of bacterial growth in vivo against Staphylococcus aureus penetrations during implantation surgery (immediate infection) and even 4 weeks after implantation (delayed infection). Thus, our antibiotic-loaded MAP hydrogel coating can open new avenues for self-defensive antibiotic prophylaxis to achieve instant and sustainable bacteriocidal activity in orthopedic prostheses.© 2017 Elsevier Inc. All rights reserved.

Sensitivity of Botrytis cinerea isolate collected from gladiolus against selected fungicides, plant oils and botanicals in North India

Integrated disease management is the best and most environmentally friendly technique for managing of a plant disease. Among the nine fungicides tested in vitro against Botrytis cinerea, which causes grey mould in Gladiolus, hexaconazole and a combination of iprodione+carbendazim exhibited complete inhibition of the mycelia growth at all the tested concentrations. In the evaluation of 15 different plant oils against B. cinerea, oils of Mentha piperita, Cymbopogon martini, Pelargonium graveolens, Cymbopogon sp., and Oreganum vulgare resulted in 100% mycelial growth inhibition of B. cinerea at all concentrations ranging from 200 to 1000 ppm. Aqueous extract of cloves of Allium sativum resulted best among the different botanicals and bioproducts tested in vitro with a maximum average mycelial growth inhibition of 57.39%, followed by the leaves of Azadirachta indica (45.47%) at 20, 50, 75, and 100 ppm. In integrated management of the grey mould of gladiolus under in vivo conditions, the combination of quintal + A. sativum + neem oil exhibited the maximum reduction in disease (94.40 %), followed by the combination of contaf + A.sativum + neem oil, which showed 93.19% disease reduction. The treatments viz., quintal +A.sativum + neem oil also resulted in the most superior treatment in enhancing growth and yield parameters of gladiolus with significantly improved plant height (70.34 cm), yield of corms (38.00), spike length (53.17 cm), number of florets (15.65), and number of flowers (32.67). These results indicate that the integrated management approach by using fungicides, botanicals and essential oils could be used for the control of grey mould diseases caused by Botrytis pathogens.

ROS-mediated antifungal activity of Ocimum essential oil-loaded nanoemulsions against postharvest fungal pathogens of Kinnow

Kinnow fruit is prone to fungal and microbial attacks, leading to significant postharvest losses during marketing and storage due to two most prominent fungal pathogens, Penicillium digitatum and P. italicum. Safer alternatives of synthetic fungicides are required for management of these pathogens. Essential oils (EO) with remarkable antimicrobial activities may offer an upcoming alternative, but the high volatility, thermal sensitivity, and hydrophobicity limit their commercial applications. In the current study, coarse and nanoemulsions (CEs and NEs) of the most potent Ocimum species were evaluated as novel coatings for Kinnow. In a screening in vitro study, the efficacy of Ocimum essential oil of three species against P. digitatum and P. italicum was assessed. The CEs and NEs (0.5%–0.3% concentrations) of Ocimum viride EO (OVEO, best screened EO) with three surfactants in 1:1 oil-surfactant proportion (OSP) were prepared. The NE formulated with 1% oil having OSP (1:1) sonicated for 10 min exhibited consistent stability till 1 month with mean droplet diameter of 180.2 nm, polydispersity index 0.328, ζ-potential (−22.58 mV) and pH of 7.84) exhibited complete inhibition of hyphal growth of test fungi at 500 mg L−1. The EO-NE treatment of the fungal hyphae led to enhanced generation of reactive oxygen species (ROS) manifested as hyphal distortion and suppression of spore germination as compared to EO. OVEO NEs may offer an ecofriendly alternate to synthetic analogues for postharvest management of Kinnow.

Nano-emulsion of three essential oils as the potential eco-friendly control to Phytophthora capsici

Phytophthora capsici is a plant pathogenic oomycete that causes significant damage to many crops, including black peppers, chili, cucurbits, and tomatoes. This pathogen is widespread and can survive in soil and plant debris for several years, making it difficult to control. The aim of the research was to observe the effectiveness of three essential oils as nano-emulsion formulas (NS, NC, NE and NA) against the growth of P. capsici in vitro and in vivo. In vitro testing was carried out using 2 methods, namely the poison food and volatile disc assay. Testing of active ingredients using the GCMS method and measuring active ingredient particles using the DLS method. In vivo testing was carried out on chili seeds to determine their growth and germination. The result indicated that all nano-emulsions at 0.50% inhibited the growth of P. capsici which varied between 33,25% to 88,50%. The NC, NS, and NA formula caused complete mycelial growth inhibition at a concentration of 1.0%. Total inhibition of growth also occurred with volatile compounds produced by NS and NA formula. The NC, NS, NE and NA formulas are not phytotoxicity to chili seeds and their germination. Therefore, three essential oils (NC, NS, NE) and the mixture of them (NA) are potential to be used in managing P. capsici as eco-friendly control and possibly as an alternative to synthetic fungicides.

Abstract A105: Pharmacological inhibition of nicotinamide adenine dinucleotide (NAD+) production enzyme nicotinamide phosphoribosyltransferase (NAMPT) impairs cellular survival, energy metabolism, and tumor growth in neuroblastoma (NB) models

Abstract Background: Neuroblastoma is the most common extracranial solid tumor in children. High-risk patients have a poor prognosis with an overall survival rate of <50%, thus novel therapeutic strategies are critically needed. To address this need, we performed a high-throughput drug screen and identified nicotinamide phosphoribosyltransferase inhibitors (NAMPTis) as highly active against neuroblastoma cells compared to other cancer cell types. NAMPT is the rate-limiting enzyme in the nicotinamide adenine dinucleotide (NAD+) salvage pathway, which cancer cells preferentially utilize to generate NAD+, a key co-factor that plays essential roles in energy metabolism. In this study, we investigated the translational potential and mechanistic effects of NAMPT inhibition in preclinical neuroblastoma models. Methods/Results: Using 3 NAMPTis on a panel of 10 neuroblastoma cell lines, we confirmed that a majority of these models were highly responsive to NAMPTis by Incucyte Live-Cell proliferation analysis. Notably, we observed complete growth inhibition in most models at doses <3.2nM of OT-82, a clinically relevant NAMPTi with a more favorable toxicity profile. On-target activity was confirmed by dose-dependent depletion of intracellular NAD+ in cells treated with OT-82. As NAD+ is necessary for energy metabolism, particularly ATP production, we performed a Cell-Titer Glo assay to indirectly assess ATP levels and observed decreases in all models with OT-82 treatment. Glycolysis and oxidative phosphorylation (OXPHOS) are two processes involved in ATP production that utilize NAD+, thus we measured activity of both processes via biochemical and extracellular flux analyses. With OT-82 treatment, we observed decreases in glucose consumption and lactate production in most models, and decreases in OXPHOS in some cell lines. Finally, an in vivo pilot study testing OT-82 in an orthotopic neuroblastoma PDX model demonstrated antitumor activity. Conclusion: Together, these data show that neuroblastoma is susceptible to disruption of the NAMPT pathway and suggest that NAMPTis have translational potential as a novel therapy for neuroblastoma patients. Citation Format: Sophia Varriano, Unsun Lee, Victor Collins, Rosa Nguyen, Sameer Issaq, Ming Sun, Ariana Nelson, Choh Yeung, Craig Thomas, Arnulfo Mendoza, Carol Thiele, Christine Heske. Pharmacological inhibition of nicotinamide adenine dinucleotide (NAD+) production enzyme nicotinamide phosphoribosyltransferase (NAMPT) impairs cellular survival, energy metabolism, and tumor growth in neuroblastoma (NB) models [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A105.

Tomato plant extract (Lycopersicon esculentum) obtained from agroindustrial byproducts and its antifungal activity against Fusarium spp.

Phytopathogenic fungi are a constant danger in the production of different crops around the world, especially in melons, since they can cause significant economic losses during the harvest, affecting the quality and shelf life. In recent years, producers have increasingly used chemical pesticides indiscriminately, causing environmental problems and damage to public health. For this reason, phytopathogenic fungi become more resistant. However, it is essential to guarantee the safety, quality, and shelf life of food after harvest, during transportation, storage and marketing. The presence of fungi in food can cause diseases transmitted through the production of toxins. Most producers depend on the discriminated use of chemical pesticides, which is a great challenge to guarantee food safety and sustainable agricultural production. To solve this problem, some extracts derived from tomato plants after harvest containing bioactive compounds have been implemented. These compounds can be natural antifungal agents as they contain phenols, flavonoids, and vitamins. Bioactive compounds emerge as a sustainable and safe opportunity in the search for new antifungal and antimicrobial agents. Therefore, the objective of this study was to determine the in vitro antifungal activity of whole tomato plant extracts on three phytopathogenic fungi. The research findings indicated that a concentration of 74.7 μg/mL of TPE resulted in a complete inhibition of mycelial growth in Fusarium oxysporum, Fusarium graminearum, and Fusarium verticillioides. Additionally, TPE exhibited both fungistatic and fungicidal effects on these Fusarium species, with a MIC50 of 30.7, 31.5, and 29.5, and a MFC of 82.4, 78.6, and 75.8 μg/mL, respectively. As a result, this study suggests that TPE can be considered as an environmentally friendly solution for extracting tomato plants, which can be applied to the surface of whole fruits or incorporated into semi-processed foods.

New Steroidal Selenides as Proapoptotic Factors.

Cytostatic and pro-apoptotic effects of selenium steroid derivatives against HeLa cells were determined. The highest cytostatic activity was shown by derivative 4 (GI50 25.0 µM, almost complete growth inhibition after three days of culture, and over 97% of apoptotic and dead cells at 200 µM). The results of our study (cell number measurements, apoptosis profile, relative expression of apoptosis-related APAF1, BID, and mevalonate pathway-involved HMGCR, SQLE, CYP51A1, and PDHB genes, and computational chemistry data) support the hypothesis that tested selenosteroids induce the extrinsic pathway of apoptosis by affecting the cell membrane as cholesterol antimetabolites. An additional mechanism of action is possible through a direct action of derivative 4 to inhibit PDHB expression in a way similar to steroid hormones.

Photodynamic inactivation by hypericin-P123 on azole-resistant isolates of the Trichophyton rubrum complex as planktonic cells and biofilm

IntroductionThe Trichophyton rubrum complex comprises the majority of dermatophyte fungi (DM) responsible for chronic cases of onychomycosis, which is treated with oral or topical antifungals. However, owing to antifungal resistance, alternative therapies, such as photodynamic therapy (PDT), are needed. This study investigated the frequency of the T. rubrum species complex in onychomycosis cases in the northwestern region of Paraná state, Brazil, and evaluated the efficacy of (PDT) using P123-encapsulated hypericin (Hyp-P123) on clinical isolates of T. rubrum in the planktonic cell and biofilm forms. Material and methodsThe frequency of the T. rubrum complex in onychomycosis cases from 2017 to 2021 was evaluated through a data survey of records from the Laboratory of Medical Mycology (LEPAC) of the State University of Maringa (UEM). To determine the effect of PDT-Hyp-P123 on planktonic cells of T. rubrum isolates, 1 × 105 conidia/mL were treated with ten different concentrations of Hyp-P123 and then irradiated with 37.8 J/cm2. Antibiofilm activity of PDT-Hyp-P123 was tested against T. rubrum biofilm in the adhesion phase (3 h), evaluated 72 h after irradiation (37.8 J/cm2), and the mature biofilm (72 h), evaluated immediately after irradiation. In this context, three different parameters were evaluated: cell viability, metabolic activity and total biomass. ResultsThe T. rubrum species complex was the most frequently isolated DM in onychomycosis cases (approximately 80 %). A significant reduction in fungal growth was observed for 75 % of the clinical isolates tested with a concentration from 0.19 μmol/L Hyp-P123, and 56.25 % had complete inhibition of fungal growth (fungicidal action); while all isolates were azole-resistant. The biofilm of T. rubrum isolates (TR0022 and TR0870) was inactivated in both the adhesion phase and the mature biofilm. ConclusionPDT-Hyp-P123 had antifungal and antibiofilm activity on T. rubrum, which is an important dermatophyte responsible for onychomycosis cases.

Efficacy of Lemon Myrtle Essential Oil as a Bio-Fungicide in Inhibiting Citrus Green Mould.

The effectiveness of lemon myrtle (LM) (Backhousia citriodora) essential oil (EO) was investigated to combat Penicillium digitatum by in vitro agar diffusion and vapour assay and in artificially infected oranges. The main constituent of LM EO was revealed as citral when analysed in gas chromatography-mass spectrometry. Pure citral was also included in the experiment for comparison. The in vitro fungal growth was significantly inhibited by LM EO at 1, 2, 3, 4 and 5 μL per disc while complete growth inhibition by both the pure citral and LM EO occurred at 4 and 5 μL per disc. Inoculated fruits treated by dipping in 1000 μL L-1 LM EO solutions for 5, 10, 15, 30 and 120 s showed significantly lower fungal wounds compared to control. While longer dipping times led to some rind injuries, fruits with a 5 and 10 s dip were found free from any injury. The evaluation after dipping and storage confirmed that the fruits maintained the sensory attributes and were not compromised by the incorporation of the essential oil. The results of this study indicate that LM EO can be a promising alternative to synthetic fungicides for preserving the quality of citrus fruits during storage.

Enhanced synergy of pacritinib with temsirolimus and sunitinib in preclinical renal cell carcinoma model by targeting JAK2/STAT pathway

Pacritinib is an oral medication that inhibits several kinases including JAK2, FLT3, IRAK and STAT3. It has been recently approved to treat patients with thrombocytopenia and myelofibrosis. Studies are currently exploring the potential use of pacritinib in treating other types of cancer such as leukaemia, breast cancer and prostate cancer. Our study aimed to investigate the effects of pacritinib alone and its combination with standard of care in renal cell carcinoma (RCC). We showed that pacritinib dose-dependently decreased viability of RCC cells, with IC50 at nanomolar or low micromolar concentration rage. Pacritinib inhibited cell proliferation, decreased colony formation, and increased apoptosis. Interestingly, pacritinib exhibited synergistic effects when combined with temsirolimus and sunitinib, but antagonistic effects when combined with doxorubicin, in a panel of RCC cell lines. We also confirmed that the combination of pacritinib with temsirolimus and sunitinib resulted in synergistic effects in RCC mouse models, with complete inhibition of tumour growth throughout the treatment period. Mechanistic studies indicated that the inhibition of JAK2, but not IRAK, was the main contributor to the anti-RCC activity of pacritinib. Our study is the first to demonstrate that pacritinib shows promise as a treatment option for RCC and underscores the therapeutic potential of targeting the JAK2/STAT signalling pathway in RCC.